Abstract
Surrogate genetically encoded markers have been utilized in order to analyze gene transfer efficacy, location, and persistence. These marker genes have greatly accelerated the development of gene transfer vectors for the ultimate application of gene therapy using therapeutic genes. They have also been used in many other applications, such as gene marking in order to study developmental cell lineages, to track cell migration, and to study tumor growth and metastasis. This chapter aims to describe the analysis of several commonly used marker genes: green fluorescent protein (GFP), β-galactosidase, firefly luciferase, human factor IX, and alkaline phosphatase. The merits and disadvantages of each are briefly discussed. In addition a few short examples are provided for continual and endpoint analysis in different disease models including hemophilia, cystic fibrosis, ornithine transcarbamylase deficiency and Gaucher disease.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Eglitis MA, Kantoff P, Gilboa E et al (1985) Gene expression in mice after high efficiency retroviral-mediated gene transfer. Science 230:1395–1398
Morral N, O’Neal W, Zhou H et al (1997) Immune responses to reporter proteins and high viral dose limit duration of expression with adenoviral vectors: comparison of E2a wild type and E2a deleted vectors. Hum Gene Ther 8:1275–1286
Berger J, Hauber J, Hauber R et al (1988) Secreted placental alkaline phosphatase: a powerful new quantitative indicator of gene expression in eukaryotic cells. Gene 66:1–10
Christou C, Parks RJ (2011) Rational design of murine secreted alkaline phosphatase for enhanced performance as a reporter gene in mouse gene therapy preclinical studies. Hum Gene Ther 22:499–506
Naylor LH (1999) Reporter gene technology: the future looks bright. Biochem Pharmacol 58:749–757
Waddington SN, Nivsarkar M, Mistry A et al (2004) Permanent phenotypic correction of Haemophilia B in immunocompetent mice by prenatal gene therapy. Blood 104:2714–2721
Misteli T, Spector DL (1997) Applications of the green fluorescent protein in cell biology and biotechnology. Nat Biotechnol 15:961–964
Wu JC, Sundaresan G, Iyer M et al (2001) Noninvasive optical imaging of firefly luciferase reporter gene expression in skeletal muscles of living mice. Mol Ther 4:297–306
Buckley SMK, Howe SJ, Rahim AA et al (2008) Luciferin detection after intra-nasal vector delivery is improved by intra-nasal rather than intra-peritoneal luciferin administration. Hum Gene Ther 19:1050–1056
Branchini BR, Ablamsky DM, Davis AL et al (2010) Red-emitting luciferases for bioluminescence reporter and imaging applications. Anal Biochem 396:290–297
Branchini BR, Ablamsky DM, Murtiashaw MH, et al (2006) Thermostable red and green light-producing firefly luciferase mutants for bioluminescent reporter applications. Anal Biochem 22:499–506
Liu HS, Jan MS, Chou CK et al (1999) Is green fluorescent protein toxic to the living cells? Biochem Biophys Res Commun 260:712–717
Bell P, Vandenberghe LH, Wu D et al (2007) A comparative analysis of novel fluorescent proteins as reporters for gene transfer studies. J Histochem Cytochem 55:931–939
Mian A, McCormack WM Jr, Mane V et al (2004) Long-term correction of ornithine transcarbamylase deficiency by WPRE-mediated overexpression using a helper-dependent adenovirus. Mol Ther 10:492–499
Enquist IB, Bianco CL, Ooka A et al (2007) Murine models of acute neuronopathic Gaucher disease. Proc Natl Acad Sci U S A 104: 17483–17488
Franks NP, Jenkins A, Conti E et al (1998) Structural basis for the inhibition of firefly luciferase by a general anesthetic. Biophys J 75: 2205–2211
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC.
About this protocol
Cite this protocol
Delhove, J.M.K.M., Rahim, A.A., McKay, T.R., Waddington, S.N., Buckley, S.M.K. (2012). Choice of Surrogate and Physiological Markers for Prenatal Gene Therapy. In: Coutelle, C., Waddington, S. (eds) Prenatal Gene Therapy. Methods in Molecular Biology, vol 891. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-873-3_13
Download citation
DOI: https://doi.org/10.1007/978-1-61779-873-3_13
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-872-6
Online ISBN: 978-1-61779-873-3
eBook Packages: Springer Protocols